EP0304150B1 - Milieu de culture sans sérum - Google Patents
Milieu de culture sans sérum Download PDFInfo
- Publication number
- EP0304150B1 EP0304150B1 EP88306022A EP88306022A EP0304150B1 EP 0304150 B1 EP0304150 B1 EP 0304150B1 EP 88306022 A EP88306022 A EP 88306022A EP 88306022 A EP88306022 A EP 88306022A EP 0304150 B1 EP0304150 B1 EP 0304150B1
- Authority
- EP
- European Patent Office
- Prior art keywords
- human
- serum
- medium
- free
- retinoic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
- C12N5/12—Fused cells, e.g. hybridomas
- C12N5/16—Animal cells
- C12N5/163—Animal cells one of the fusion partners being a B or a T lymphocyte
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
- C12N5/12—Fused cells, e.g. hybridomas
- C12N5/16—Animal cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/05—Inorganic components
- C12N2500/10—Metals; Metal chelators
- C12N2500/20—Transition metals
- C12N2500/24—Iron; Fe chelators; Transferrin
- C12N2500/25—Insulin-transferrin; Insulin-transferrin-selenium
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/38—Hormones with nuclear receptors
- C12N2501/385—Hormones with nuclear receptors of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/863—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof involving IgM
- Y10S530/864—Monoclonal
- Y10S530/865—Human
Definitions
- This invention relates to a process for cultivating a human monoclonal antibody-producing human/human hybridoma, for example in the analysis of biochemical mechanisms and the production of bio-products useful for diagnostic, prophylactic and therapeutic purposes in the field of biotechnology. More specifically, it relates to a process for cultivating a human monoclonal antibody-producing human/human hybridoma in a serum-free medium, in which the human/human hybridoma can be cultivated to give a human monoclonal antibody in a markedly improved output and subcultivation suitable for industrial practice can be carried out.
- Serum contains a variety of foreign heterogeneous factors including high-density lipolipid (HDL) and low density lipolipid (LDL), and the inclusion of such undesirable heterogenous factors in the media cannot be avoided.
- HDL high-density lipolipid
- LDL low density lipolipid
- the use of serum-containing complete media in the analysis of biochemical mechanisms such as the cell proliferation mechanism and the antibody production mechanism or in the production of bio-products such as antibodies or other useful substances causes various technical troubles. For example, the analysis of bio-chemical mechanisms is impeded, or highly pure useful bio-substances of high quality free from inclusion of heterogeneous factors from the serum added to the media cannot be obtained easily.
- the use of the serum-free complete medium containing retinoic acid or a salt thereof markedly increases the antibody-producing ability of the human-human hybridoma, and makes it possible to perform sub-cultivation of the hybridoma smoothly on an industrial scale.
- a serum-free complete medium composed of a basal medium for animal cell cultivation and at least one growth factor selected from insulin and transferrin and being free from serum is used.
- retinoic acid in a concentration of at least 10 ⁇ 12 M but not more than 10 ⁇ 6 M, preferably at least 10 ⁇ 10 M but not more than 10 ⁇ 6 M, more preferably at least 10 ⁇ 9 M but not more than 10 ⁇ 7 M in this serum-free complete medium, a serum free medium for culturing a human monoclonal antibody-producing human/human hybridoma is provided.
- a serum-free medium composed essentially of the above components (1), (2), (4), (5) and (6), a serum-free medium composed essentially of the above components (1), (2), (4), (5) and (7), a serum-free medium composed essentially of the above components (1), (2), (4) and (5), and a serum-free medium composed essentially of the above components (1), (2) and (5) may also be used in this invention by including a specific amount of retinoic acid or its salt.
- basal medium (1) for animal cell culture are known and can be prepared in accordance with the known literature (for example, "Cell Culture Manual”, 3rd edition, July 20, 1984, published by Kodansha Scientific). Many of them are commercially available, and can be utilized in this invention.
- basal medium examples include known basal media and known modified media thereof shown below.
- BME medium Basal Medium Eagle
- Eagle described, for example, in Eagle, H.: Science, 122, 501 (1955); Eagle, H.: J. Exp. Med., 102, 37(1955) and 102, 595 (1955); Eagle, H.: J. Biol. Chem., 214, 839 (1955); Eagle, H. et al.: Science, 123, 845 (1955); Hanks, J. H., Wallace, R. E.: Proc. Soc. Exp. Biol. Med., 71, 196 (1949); Yamane, I.: Proc. Soc. Exp. Biol. Med., 127, 335 (1968); Morton, H. J.: In Vitro, 6, 89 (1970); and Eagle, H.: Proc. Soc. Exp. Biol. Med., 89, 362 (1955).
- MEM medium Minimum Essential Medium described, for example, in Eagle, H.: Science, 130, 432 (1959); Stoker, M., MacPherson, I.: Virology, 14, 359 (1931); MacPherson, I., Stoker, M.: Virology, 16, 147 (1962); Stoker, M., MacPherson, I.: Nature, 203, 1355 (1964); Dulbecco, R., Freeman, G.: Virology, 8, 396 (1959); Smith J. D., et al.: Virology, 12, 185 (1960); Stanners, C. P. et al.: Nature New Biology, 230, 52 (1971); and Stanners, C. P., Stewart, C.: Personal Communication (1972).
- Williams' Medium E described, for example, in Williams, G. M., Weisburger, E. K. and Weisburger, J. H.: Exp. Cell Res., 69, 106-112 (1971).
- NCTC 135 Medium described, for example, in Evans, V. J. et al.: Exp. Cell Res., 36, 439 (1968).
- Waymouth's Medium MB752/1 described, for example, in Waymouth, C: J. Nat. Cancer Inst., 22, 1003 (1959), and Morton, H. J.: In Vitro, 6, 89 (1970).
- basal media for animal cell culture may be used singly or as mixtures in suitable proportions.
- the serum-free medium used in cultivating a human monoclonal antibody-producing human/human hybridoma is composed of such a serum-free complete medium as illustrated above and at least 10 ⁇ 12 M but not more than 10 ⁇ 6 M of retinoic acid or its salt.
- the amount of retinoic acid or its salt may be varied within the above range depending upon various factors such as the types of the basal medium constituting the serum-free complete medium and the complete medium-forming additives, their combinations and proportions, the type of the human/human hybridoma to be cultivated and the purpose of cultivation. Those skilled in the art, if required, can select and determine preferred amounts of retinoic acid or its salt according to these factors.
- the preferred amount is, for example, at least 10 ⁇ 10 M but not more than 10 ⁇ 6 M, especially at least 10 ⁇ 9 M but not more than 10 ⁇ 7 M.
- the especially preferred amount of retinoic acid or its salt is, for example, at least 10 ⁇ 9 M but less than 10 ⁇ 7 M.
- the output of the antibody tends to decrease considerably if the content of retinoic acid or its salt is smaller or larger beyond the above-specified range.
- the human/human hybridoma and the method of its formation are not the subject matter of this invention.
- Some examples of such hybridomas include the human monoclonal antibody-producing human/human hybridomas disclosed in Japanese Laid-Open Patent Publications Nos. 201994/1983, 13589/1984, 137497/1984, and 70400/1987 which can be obtained by the methods disclosed in these patent documents; and the human monoclonal antibody-producing human/human hybridomas disclosed in detail in Japanese Laid-Open Patent Publication No. 155083/1987 and Japanese Patent Applications Nos. 202752/1986 filed on August 30, 1986, and 126687/1987 filed on May 23, 1987 which can be obtained by utilizing the techniques disclosed in the first four Japanese Laid-Open Patent Publications cited above.
- human/human hybridomas are human/human hybridoma CLN/SUZH5 (Fermentation Research Institute, Deposition Rejecting Notice No. 57-637), human/human hybridoma CLN H5 (ATCC HB8206) and human/human hybridoma SLN F10 (Fermentation Research Institute, Deposition Rejecting Notice No. 60-1197), which are disclosed in the above-cited patent documents; human/human hybridoma CoLNE10 (Fermentation Research Institute, Deposition Rejecting Notice No.
- fusion partners typified as HIH/TOl given as a typical example of fusion partner above and the techniques of forming human/human hybridomas using them will be described below briefly although they are not the subject matter of the present invention.
- the above fusion partner can be obtained from human B cell lymphoblast cells W1-L2 (Fermentation Research Institute, Deposition Rejecting Notice No. 60-1621) by a mutant forming operation in accordance with a technique of making it drug-resistant.
- the resulting cell line having self-replicability is a mutant derived from human B cell lymphoblast cells and has the following characteristics (i) to (v).
- the above fusion partner which is a human B cell lymphoblast cell mutant can be created by, for example, cultivating and adapting human B cell lymphoblast cells in a serum-free medium, screening the adapted cells to select cells substantially lacking the ability to product immunoglobulins, cultivating and adapting the selected cells in a 6-thioguanine-containing serum-free medium, treating the resulting 6-thioguanin-resistant cells with a mutating agent, cultivating and adapting the treated cells in an ouabain-containing serum-free medium, and cloning the resistant cells in a serum-free medium containing both 6-thioguanine and ouabain.
- a preferred example of the serum-free medium used at this time is a serum-free medium prepared by adding suitable amounts of insulin, transferrin, selenium, ethanolamine, ⁇ -mercaptoethanol and bovine serum albumin to basal medium RDF (a mixture of RPMI 1640, DME and F12 in a ratio of 2:1:1).
- basal medium RDF a mixture of RPMI 1640, DME and F12 in a ratio of 2:1:1.
- Other serum-free media may also be used, and they may be experimentally selected, changed or determined according to the types of the human B cell lymphoblast cells to be cultivated and the basal medium, the types and amounts of the additives to the basal medium, etc.
- mutating agent examples include known ones such as MNNG, EMS, AAB, AAF, AF-2, BP 0x , DAB, BZD, DAN, DBA, DBE, DBP, DMN, ENNG, ENU, HFA, 3MCA, MMS, 2NA, NAAAF, NBA, 4NQO, OAT, PI, TCE, TDS, TOX and VC.
- Colonies where the presence of fused cells are observed are selected, and examined for a human immuloglobulin by, for example, radioimmunoassay using 125I, or an enzyme-linked immunosorbent assay.
- the selected colonies where the production of the human immunoglobulin is determined are transferred to a fresh culture medium and cultivated to proliferate the fused cells and thus obtain fused cell clones.
- the serum-free culture medium containing a specific amount of retinoic acid or its salt is utilized for cultivating human monoclonal antibody-producing human/human hybridomas.
- the serum-free medium may contain a predetermined amount of retinoic acid.
- retinoic acid may be added to the medium at the time of use so that its amount may reach the above predetermined amount.
- the cultivation conditions may be properly selected, and if required, may be changed by performing a preliminary experiment.
- the cultivation may be carried out at about 37 ⁇ 3 °C in an atmosphere containing about 5 % of CO2.
- the following Examples illustrate the preparation of the serum-free medium and the cultivation of a human monoclonal antibody-producing human/human hybridoma in the serum-free medium.
- Example 1 The number of cells and the amount of the antibody are shown in Table 1.
- Figure 1 shows the relation between the amount of the antibody and the amount of retinoic acid.
- Table 1 Run Amount of retinoic acid in the serum-free complete medium (M) Number of cells 6 days after start of cultivation (x 106 cells/ml) Amount of the antibody 6 days after start of cultivation ( ⁇ g/ml) Control 0 2.75 1
- Example 1 10 ⁇ 10 - 3.2
- Example 2 10 ⁇ 9 3.2 3.7
- Example 3 10 ⁇ 8 - 3.9
- Example 4 10 ⁇ 7 3.06 3.8
- Example 5 10 ⁇ 6 - 3.5 Comparative Example 1 10 ⁇ 5 0.22 (below the detection limit) -: Not measured
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Immunology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Claims (3)
- Procédé pour cultiver un hybridome humain/humain producteur d'anticorps monoclonaux humains, dans lequel cet hybridome est cultivé dans un milieu complet sans sérum contenant au moins 10⁻¹² M, mais pas plus de 10⁻⁶ M d'acide rétinoïque ou d'un sel de celui-ci.
- Procédé selon la revendication 1, dans lequel la quantité d'acide rétinoïque est 10⁻⁹ M, mais pas plus de 10⁻⁷ M.
- Procédé selon la revendication 1, dans lequel ce milieu complet sans sérum comprend un milieu de base pour la culture de cellules animales et, comme facteur de croissance, l'insuline et/ou la transferrine.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP62206569A JPH0634713B2 (ja) | 1987-08-21 | 1987-08-21 | ヒト/ヒト・ハイブリドーマの培養方法 |
JP206569/87 | 1987-08-21 |
Publications (3)
Publication Number | Publication Date |
---|---|
EP0304150A2 EP0304150A2 (fr) | 1989-02-22 |
EP0304150A3 EP0304150A3 (en) | 1989-08-30 |
EP0304150B1 true EP0304150B1 (fr) | 1994-01-12 |
Family
ID=16525567
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP88306022A Expired - Lifetime EP0304150B1 (fr) | 1987-08-21 | 1988-07-01 | Milieu de culture sans sérum |
Country Status (10)
Country | Link |
---|---|
US (1) | US5155036A (fr) |
EP (1) | EP0304150B1 (fr) |
JP (1) | JPH0634713B2 (fr) |
KR (1) | KR940003651B1 (fr) |
AU (1) | AU615918B2 (fr) |
CA (1) | CA1299509C (fr) |
DE (1) | DE3887030T2 (fr) |
ES (1) | ES2061655T3 (fr) |
IL (1) | IL86830A0 (fr) |
RU (1) | RU2018532C1 (fr) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107099508A (zh) * | 2017-06-23 | 2017-08-29 | 曲宝赤 | 一种无血清杂交瘤细胞培养基 |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2123094C (fr) * | 1991-11-06 | 1999-08-10 | Paulo N. Correa | Milieu de culture cellulaire |
US6037174A (en) * | 1993-08-23 | 2000-03-14 | Nexell Therapeutics, Inc. | Preparation of serum-free suspensions of human hematopoietic cells or precursor cells |
US5846529A (en) * | 1993-08-23 | 1998-12-08 | Nexell Therapeutics, Inc. | Infusion of neutrophil precursors for treatment of neutropenia |
US20030077757A1 (en) * | 2000-01-11 | 2003-04-24 | Andrews William H. | Method of treating aging-related disorders |
WO2004029069A2 (fr) * | 2002-09-30 | 2004-04-08 | Pfizer Products Inc. | Hybridomes generant des taux eleves d'anticorps contre des sequences humaines |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4205126A (en) * | 1978-01-01 | 1980-05-27 | Cartaya Oscar A | Serum-free cell culture media |
-
1987
- 1987-08-21 JP JP62206569A patent/JPH0634713B2/ja not_active Expired - Fee Related
-
1988
- 1988-06-22 IL IL86830A patent/IL86830A0/xx not_active IP Right Cessation
- 1988-06-27 AU AU18421/88A patent/AU615918B2/en not_active Ceased
- 1988-07-01 ES ES88306022T patent/ES2061655T3/es not_active Expired - Lifetime
- 1988-07-01 EP EP88306022A patent/EP0304150B1/fr not_active Expired - Lifetime
- 1988-07-01 DE DE88306022T patent/DE3887030T2/de not_active Expired - Fee Related
- 1988-08-11 US US07/230,973 patent/US5155036A/en not_active Expired - Lifetime
- 1988-08-17 CA CA000574947A patent/CA1299509C/fr not_active Expired - Lifetime
- 1988-08-19 RU SU884356208A patent/RU2018532C1/ru not_active IP Right Cessation
- 1988-08-20 KR KR1019880010575A patent/KR940003651B1/ko not_active IP Right Cessation
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107099508A (zh) * | 2017-06-23 | 2017-08-29 | 曲宝赤 | 一种无血清杂交瘤细胞培养基 |
Also Published As
Publication number | Publication date |
---|---|
KR940003651B1 (ko) | 1994-04-25 |
AU615918B2 (en) | 1991-10-17 |
DE3887030D1 (de) | 1994-02-24 |
RU2018532C1 (ru) | 1994-08-30 |
US5155036A (en) | 1992-10-13 |
JPS6451079A (en) | 1989-02-27 |
ES2061655T3 (es) | 1994-12-16 |
AU1842188A (en) | 1989-02-23 |
IL86830A0 (en) | 1988-11-30 |
JPH0634713B2 (ja) | 1994-05-11 |
KR890003944A (ko) | 1989-04-19 |
DE3887030T2 (de) | 1994-04-28 |
EP0304150A2 (fr) | 1989-02-22 |
CA1299509C (fr) | 1992-04-28 |
EP0304150A3 (en) | 1989-08-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Glassy et al. | Serum‐free media in hybridoma culture and monoclonal antibody production | |
US5681718A (en) | Methods for enhanced production of tissue plasminogen activator in cell culture using alkanoic acids or salts thereof | |
US4594325A (en) | High fusion frequency fusible lymphoblastoid cell line | |
US4624921A (en) | Human lymphoblastold cell line and hybridomas derived therefrom | |
Shacter | Serum-free medium for growth factor-dependent and-independent plasmacytomas and hybridomas | |
Strike et al. | Production of human-human hybridomas secreting antibody to sheep erythrocytes after in vitro immunization. | |
EP0239292B1 (fr) | Production de protéines par culture de cellules | |
EP0304150B1 (fr) | Milieu de culture sans sérum | |
EP0293155B1 (fr) | Immunoglobulines humaines spécifiques de l'antigène relatif au cancer et hybridomes humains-humains pouvant produire ces immunoglobulines humaines | |
EP0076647A2 (fr) | Milieux de culture pour cellules issues du système immun | |
EP0057107A2 (fr) | Procédé de préparation d'anticorps monoclonaux et cellules aptes à produire ces anticorps | |
EP0248656B1 (fr) | Composition pour la culture de cellules et son utilisation | |
US4757018A (en) | Myeloma cell lines and uses thereof | |
CA1218947A (fr) | Lignees de cellules hybrides humaines | |
Ozturk | Kinetic characterization of hybridoma growth, metabolism, and monoclonal antibody production rates | |
Bols et al. | Media for hybridoma growth and monoclonal antibody production | |
JP2509191B2 (ja) | ガン関連抗原特異的ヒト免疫グロブリン | |
JPS60204727A (ja) | 抗ヒトv型コラ−ゲン抗体 | |
US4897466A (en) | Human lymphoblastoid cell line and hybridomas derived therefrom | |
EP0368662A2 (fr) | Lignes cellulaires parentales pour la production d'hybridomes humains | |
Darfler | In Vitro Immunization for the Generation of Hybridomas Using Serum-Free Medium | |
JPH084518B2 (ja) | モノクロナル抗体生産性ヒト/ヒトハイブリド−マの無血清培養法 | |
Lee et al. | Stability of Chimeric Antibody Producing Transfectomas during a Long-Term Culture | |
Morrill | Characterization of the down regulation of antibody production in high cell density perfusion culture | |
McKearn et al. | Mapping of Novel B Lymphocyte Differentiation Antigens |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
AK | Designated contracting states |
Kind code of ref document: A2 Designated state(s): BE CH DE ES FR GB IT LI NL SE |
|
PUAL | Search report despatched |
Free format text: ORIGINAL CODE: 0009013 |
|
AK | Designated contracting states |
Kind code of ref document: A3 Designated state(s): BE CH DE ES FR GB IT LI NL SE |
|
17P | Request for examination filed |
Effective date: 19891017 |
|
17Q | First examination report despatched |
Effective date: 19920724 |
|
GRAA | (expected) grant |
Free format text: ORIGINAL CODE: 0009210 |
|
ITF | It: translation for a ep patent filed |
Owner name: ST. DR. CAVATTONI ING. A. RAIMONDI |
|
AK | Designated contracting states |
Kind code of ref document: B1 Designated state(s): BE CH DE ES FR GB IT LI NL SE |
|
REF | Corresponds to: |
Ref document number: 3887030 Country of ref document: DE Date of ref document: 19940224 |
|
ET | Fr: translation filed | ||
PLBE | No opposition filed within time limit |
Free format text: ORIGINAL CODE: 0009261 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT |
|
REG | Reference to a national code |
Ref country code: ES Ref legal event code: FG2A Ref document number: 2061655 Country of ref document: ES Kind code of ref document: T3 |
|
26N | No opposition filed | ||
EAL | Se: european patent in force in sweden |
Ref document number: 88306022.0 |
|
REG | Reference to a national code |
Ref country code: GB Ref legal event code: IF02 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: CH Payment date: 20040629 Year of fee payment: 17 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: GB Payment date: 20040630 Year of fee payment: 17 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: NL Payment date: 20040704 Year of fee payment: 17 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: SE Payment date: 20040706 Year of fee payment: 17 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: FR Payment date: 20040708 Year of fee payment: 17 Ref country code: DE Payment date: 20040708 Year of fee payment: 17 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: ES Payment date: 20040719 Year of fee payment: 17 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: BE Payment date: 20040909 Year of fee payment: 17 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: IT Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES;WARNING: LAPSES OF ITALIAN PATENTS WITH EFFECTIVE DATE BEFORE 2007 MAY HAVE OCCURRED AT ANY TIME BEFORE 2007. THE CORRECT EFFECTIVE DATE MAY BE DIFFERENT FROM THE ONE RECORDED. Effective date: 20050701 Ref country code: GB Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20050701 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: SE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20050702 Ref country code: ES Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20050702 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: LI Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20050731 Ref country code: CH Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20050731 Ref country code: BE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20050731 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: NL Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20060201 Ref country code: DE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20060201 |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: PL |
|
EUG | Se: european patent has lapsed | ||
GBPC | Gb: european patent ceased through non-payment of renewal fee |
Effective date: 20050701 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: FR Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20060331 |
|
NLV4 | Nl: lapsed or anulled due to non-payment of the annual fee |
Effective date: 20060201 |
|
REG | Reference to a national code |
Ref country code: FR Ref legal event code: ST Effective date: 20060331 |
|
REG | Reference to a national code |
Ref country code: ES Ref legal event code: FD2A Effective date: 20050702 |
|
BERE | Be: lapsed |
Owner name: *HAGIWARA HIDEAKI Effective date: 20050731 Owner name: *HAGIWARA YOSHIHIDE Effective date: 20050731 |